Precise studies of plant, animal and human genomes enable remarkable
opportunities of obtained data application in biotechnology and medicine.
However, knowing nucleotide sequences isn’t enough for understanding of
particular genomic elements functional relationship and their role in phenotype
formation and disease pathogenesis. In post-genomic era methods allowing
genomic DNA sequences manipulation, visualization and regulation of gene
expression are rapidly evolving. Though, there are few methods, that meet high
standards of efficiency, safety and accessibility for a wide range of
researchers. In 2011 and 2013 novel methods of genome editing appeared –
this are TALEN (Transcription Activator-Like Effector Nucleases) and CRISPR
(Clustered Regulatory Interspaced Short Palindromic Repeats)/Cas9 systems.
Although TALEN and CRISPR/Cas9 appeared recently, these systems have proved to
be effective and reliable tools for genome engineering. Here we generally
review application of these systems for genome editing in conventional model
objects of current biology, functional genome screening, cell-based human
hereditary disease modeling, epigenome studies and visualization of cellular
processes. Additionally, we review general strategies for designing TALEN and
CRISPR/Cas9 and analyzing their activity. We also discuss some obstacles
researcher can face using these genome editing tools.
Induced pluripotent stem cells (iPSCs) are a new type of pluripotent cells
that can be obtained by reprogramming animal and human differentiated cells. In this review,
issues related to the nature of iPSCs are discussed and different methods of
iPSC production are described. We particularly focused on methods of iPSC production without
the genetic modification of the cell genome and with means for increasing the iPSC production
efficiency. The possibility and issues related to the safety of iPSC use in cell replacement
therapy of human diseases and a study of new medicines are considered.
To date biomedicine and pharmacology have required generating new and more
consummate models. One of the most perspective trends in this field is using
induced pluripotent stem cells (iPSCs). iPSC application requires careful
high-throughput analysis at the molecular, epigenetic, and functional levels.
The methods used have revealed that the expression pattern of genes and
microRNA, DNA methylation, as well as the set and pattern of covalent histone
modifications in iPSCs, are very similar to those in embryonic stem cells.
Nevertheless, iPSCs have been shown to possess some specific features that can
be acquired during the reprogramming process or are remnants of epigenomes and
transcriptomes of the donor tissue. These residual signatures of epigenomes and
transcriptomes of the somatic tissue of origin were termed “epigenetic
memory.” In this review, we discuss the “epigenetic memory”
phenomenon in the context of the reprogramming process, its influence on iPSC
properties, and the possibilities of its application in cell technologies.
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